Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies

Paul W Tillberg, Fei Chen, Kiryl D Piatkevich, Yongxin Zhao, Chih-Chieh Jay Yu, Brian P English, Linyi Gao, Anthony Martorell, Ho-Jun Suk, Fumiaki Yoshida, Ellen M DeGennaro, Douglas H Roossien, Guanyu Gong, Uthpala Seneviratne, Steven R Tannenbaum, Robert Desimone, Dawen Cai, Edward S Boyden. Nature biotechnology, Jul 2016.


Expansion microscopy (ExM) enables imaging of preserved specimens with nanoscale precision on diffraction-limited instead of specialized super-resolution microscopes. ExM works by physically separating fluorescent probes after anchoring them to a swellable gel.

The first ExM method did not result in the retention of native proteins in the gel and relied on custom-made reagents that are not widely available. Here we describe protein retention ExM (proExM), a variant of ExM in which proteins are…